WO2019206159A1

WO2019206159A1 - Use of chlorogenic acid and composition thereof in preparation of medicament for treating sarcoma

Info

Publication number: WO2019206159A1
Application number: PCT/CN2019/083989
Authority: WO
Inventors: 张洁; 陈晓光; 黄望
Original assignee: 四川九章生物科技有限公司
Priority date: 2018-04-24
Filing date: 2019-04-24
Publication date: 2019-10-31
Also published as: US20210338622A1; CN108653263B; CN108653263A

Abstract

Disclosed in the present invention is the use of chlorogenic acid in preparation of a medicament for treating sarcoma. The present invention further provides a chlorogenic acid combination and the use thereof in preparation of a medicament for treating sarcoma. The chlorogenic acid of the present invention can effectively treat sarcoma, and the therapeutic effect is superior to that of the chemotherapeutic medicament doxorubicin when being administered systematically or topically. The chlorogenic acid composition of the present invention comprises chlorogenic acid and coumaroyl quinic acid, can effectively treat sarcoma, and has a better therapeutic effect than the chemotherapeutic medicament doxorubicin. The experimental results of the present invention show that the combination of chlorogenic acid and coumaroylquinic acid can have a synergistic effect.

Description

Use of chlorogenic acid and its composition for preparing medicine for treating sarcoma

Technical field

The invention particularly relates to the use of chlorogenic acid for the manufacture of a medicament for the treatment of sarcoma; the invention also relates to the use of a chlorogenic acid composition for the preparation of a medicament for the treatment of sarcoma.

Background technique

Sarcoma refers to malignant tumors derived from mesenchymal tissue (including connective tissue and muscle), which occur mostly at the skin, subcutaneous, periosteum and long bones. Osteosarcoma is more common in young people, and it occurs at the two ends of the long bones of the extremities, especially at the lower end of the femur, the upper end of the humerus and the upper end of the tibia. Osteosarcoma develops rapidly and has a short course of disease. It begins to grow in the cortex and can gradually develop into the medullary cavity. Sometimes it breaks through the periosteum and invades the surrounding soft tissue. It is easy to cause pathological fractures. Commonly, leiomyomas, lymphosarcoma, and synovium In sarcoma, blood transfer can occur early. Sarcoma is a malignant tumor. ,

At present, the main treatment methods for sarcoma are: surgery, chemotherapy and local radiotherapy. In surgery, amputation or radical surgery is the most common method, but the curative effect is still not satisfactory. About 50% of the patients still die, and the effect of partial resection is not satisfactory. At present, both domestic and foreign tend to retain most of the resection of the limb and then undergo radiotherapy. . Chemotherapy is feasible before and after chemotherapy, but the efficiency is not high, and side effects are large. Local radiotherapy may also be selective for some sarcoma patients, but the effectiveness is low, but the effect of lung metastasis is even worse.

Therefore, there is a need for a drug that can effectively treat sarcoma. No reports of chlorogenic acid in the treatment of sarcoma.

Summary of the invention

The technical solution of the present invention is to provide a new use of chlorogenic acid.

Use of the chlorogenic acid of the invention in the preparation of a medicament for treating sarcoma.

Wherein, the medicament is prepared by using an effective amount of chlorogenic acid as an active ingredient, and adding a pharmaceutically acceptable auxiliary or auxiliary component.

Wherein, the pharmaceutical preparation contains 1 to 3000 mg of chlorogenic acid per preparation unit.

Wherein, the medicinal preparation uses chlorogenic acid in a dose of 1 to 100 mg/kg.

Wherein the agent is an oral preparation or an injection.

Wherein, the sarcoma comprises osteosarcoma, fibrosarcoma, liposarcoma, rhabdomyosarcoma.

The present invention also provides a pharmaceutical composition comprising chlorogenic acid and coumaric quinic acid.

Further, the mass ratio of the chlorogenic acid to the coumaroyl quinic acid is 100: 0.01 to 0.5.

Preferably, the mass ratio of the chlorogenic acid to the coumaroyl quinic acid is 100: 0.01 to 0.1.

More preferably, the mass ratio of the chlorogenic acid to coumaroyl quinic acid is 100:0.05.

The present invention provides a method for preparing the above pharmaceutical composition, which comprises chlorogenic acid and coumaroyl quinic acid as active ingredients, and a pharmaceutically acceptable adjuvant is added to prepare a pharmaceutically acceptable pharmaceutical preparation.

Preferably, the preparation is an oral preparation or an injection preparation.

The present invention provides the use of the above pharmaceutical composition for the preparation of a medicament for the treatment of sarcoma.

The chlorogenic acid of the invention can effectively treat sarcoma, and the therapeutic effect is superior to the chemotherapeutic drug doxorubicin in systemic administration + local administration mode, and it has been proved to be a safe drug with very little side effects. It can replace traditional chemotherapy drugs for the treatment of sarcoma, and can effectively solve the problem of traditional chemotherapeutic drugs for the treatment of sarcoma with large side effects, and has good clinical application prospects.

The chlorogenic acid composition of the invention comprises chlorogenic acid and coumaric quinic acid, which can effectively treat sarcoma, and the therapeutic effect is superior to the chemotherapy drug doxorubicin. The experimental results of the present invention show that the combination of chlorogenic acid and coumaric quinic acid can exert synergistic effects.

It is apparent that various other modifications, substitutions and changes can be made in the form of the above-described embodiments of the present invention.

The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples. Any technique implemented based on the above description of the present invention is within the scope of the present invention.

DRAWINGS

Fig.1 Effect of experimental groups in experimental experiment 1 on growth curve of transplanted tumor of S180 sarcoma KM mice

Fig. 2 Effect of each experimental group in experimental group 1 on tumor inhibition rate of transplanted tumor of S180 sarcoma KM mice

Fig. 3 Effect of experimental groups in experimental experiment 2 on growth curve of transplanted tumor of S180 sarcoma KM mice

Fig. 4 Effect of each experimental group in experimental group 2 on the tumor inhibition rate of transplanted tumor of S180 sarcoma KM mice

detailed description

The raw materials and equipment used in the specific embodiments of the present invention are known products and are obtained by purchasing commercially available products.

Example 1 Formulation of Oral Formulation of Pharmaceutical Composition of the Invention

1, prescription one

1000 g of chlorogenic acid and 1 g of coumaric quinic acid.

Preparation method: chlorogenic acid and coumaric quinic acid are weighed according to the prescription, and evenly mixed, and then aseptically divided into powder.

2, prescription two

1000 g of chlorogenic acid, 5 g of coumaric quinic acid, 500 g of a filler, and 5 g of a binder.

Preparation method: chlorogenic acid, coumaric quinic acid, filler, binder, granulation, granules, granules and granules are weighed according to the prescription.

3, prescription three

1000 g of chlorogenic acid, 1 g of coumaric quinic acid, 500 g of a filler, 5 g of a binder, and 3 g of a lubricant.

Preparation method: chlorogenic acid, coumaric quinic acid, filler, binder, granules, whole granules, lubricants, tableting, tablets are obtained according to the prescription.

The above filler is one or more of mannitol, lactose, starch, microcrystalline cellulose, dextrin; the binder is sodium carboxymethylcellulose, PVP; the lubricant is magnesium stearate, talc, Micronized silica gel.

Example 2 Formulation of Injectable Formulation of Pharmaceutical Composition of the Present Invention

1, prescription one

1000 g of chlorogenic acid and 1 g of coumaric quinic acid.

Preparation method (1): chlorogenic acid and coumaric quinic acid are weighed according to the prescription, and after mixing, they are aseptically divided into powder injections.

Preparation method (2): chlorogenic acid and coumaric quinic acid are weighed according to the prescription, dissolved in water for injection, filtered and sterilized, and lyophilized to obtain a lyophilized powder injection.

2, prescription two

1000 g of chlorogenic acid, 1 g of coumaric quinic acid, 2667 g of scaffold, and 67 g of antioxidant.

Preparation method: chlorogenic acid, coumaric quinic acid, scaffold, antioxidant, weighed according to the prescription, dissolved in water for injection, filtered and sterilized, freeze-dried to obtain lyophilized powder injection.

The above scaffolding agents are mannitol, lactose, and glucose; and the antioxidants are sodium hydrogen sulfite, vitamins, glutathione, and folic acid.

The beneficial effects of the present invention will be described below by way of experimental examples:

Experimental Example 1 Inhibition of chlorogenic acid on KM mouse S180 sarcoma xenografts in vivo

1 experimental material

1) Animals

KM mice, half male and half female, weigh 17 to 24 g.

2) Cell line

The S180 cell line is a mouse sarcoma cell. Suspension growth in DH medium containing 20% calf serum, 1mmol/L glutamine and 100U·ml ^-1 penicillin, 100μg·ml ^-1 streptomycin at 37 ° C, 5% CO ₂ saturated humidity Culture in the box, change the liquid every 2 to 3 days.

3) drugs

Chlorogenic acid, doxorubicin (ADM), physiological saline.

2 experimental methods

1) Cell culture

S180 cells in the logarithmic growth phase were harvested, centrifuged at 1000 rpm for 5 minutes, washed twice, and the number of viable cells was counted by trypan blue staining. Inoculated as soon as possible in the peritoneal cavity of KM mice as a breeder.

2) In vivo experiment

The ascites was taken from 7 to 10 days. After dilution, the number of cells was adjusted to (about 1×10 ⁶ cells), and the ascites cell suspension 0.2 ml·only ^{-1 was} inoculated subcutaneously in the left forelimb of the mouse. Animals were randomly divided into groups of 10, each of which was chlorogenic acid group, ADM group (2 mg·kg ^-1 ) and negative group (NS), and administration was started on the second day after inoculation.

The chlorogenic acid systemic administration group was administered intravenously once a day at a dose of 30 mg·kg ^-1 .

The chlorogenic acid local administration group was administered subcutaneously in the vicinity of the in situ lesion, once a day, at a dose of 30 mg·kg ^-1 .

Systemic administration of chlorogenic acid + topical administration group: intravenous administration, once daily, the dose is 30 mg·kg ^-1 , subcutaneous injection around the lesion in situ, once every 5 days, the dose is 30 mg· Kg ^-1 .

ADM group: administered intraperitoneally, once daily, at a dose of 2 mg·kg ^-1 .

Negative group: administered intraperitoneally, once daily, given an equal volume of normal saline.

Tumor volume (V = 0.52 × long diameter of transplanted tumor L × shortest diameter W ^{2 of} transplanted tumor) was measured on the 5th day after inoculation, and the mice were weighed every three days. When the average tumor of the negative group was greater than 1 g (the tumor volume was about 0.5 cm ³ ), the experiment was stopped, the mice were sacrificed by cervical dislocation and weighed, the tumor was removed, and the tumor inhibition rate was calculated. The tumor inhibition rate was %=[1-(administration) Group mean tumor weight/negative group mean tumor weight)] × 100%.

3 experimental results

1) Effect of each experimental group on the growth volume of transplanted tumor of S180 sarcoma KM mice

See Table 1 and Figure 1.

Table 1 Effect of each experimental group on the growth volume of transplanted tumor of S180 sarcoma KM mice (

n=10)

Comparison of each drug group and negative group *p<0.05, **p<0.001

It can be seen from Table 1 and Figure 1:

1 chlorogenic acid systemic administration group, local administration group, systemic + local administration group, ADM positive control group significantly inhibited the growth volume of transplanted tumor of S180 sarcoma KM mice at different time points during the experiment. Compared with the negative control group, there was a statistical difference;

2 The systemic + local administration group inhibited the growth volume of transplanted tumor of S180 sarcoma KM mice better than the systemic administration group and the local administration group;

3 The systemic + local administration group inhibited the growth volume of S180 sarcoma KM mouse xenografts in comparison with the ADM positive control group.

2) Effects of experimental groups on the weight and tumor inhibition rate of transplanted tumor of S180 sarcoma KM mice

See Table 2 and Figure 2.

Table 2 Effect of experimental groups on the weight and tumor inhibition rate of transplanted tumor of S180 sarcoma KM mice

Compared with the negative group ***p<0.001

It can be seen from Table 2 and Figure 2 that the inhibition rate of S180 sarcoma KM mouse xenografts is higher than 40% in the chlorogenic acid systemic administration group, the local administration group, the systemic + local administration group, and the ADM positive control group; The chlorogenic acid system + local administration group had the highest tumor inhibition rate of 57%.

The above experiments show that chlorogenic acid has a significant inhibitory effect on mouse sarcoma, and when the mode of administration is systemic + local administration, the inhibition is optimal, and chlorogenic acid can be used for the treatment of sarcoma.

Experimental Example 2 Inhibition of chlorogenic acid composition on KM mouse S180 sarcoma xenografts in vivo

1 experimental material

1) Animals

KM mice, half male and half female, weigh 17 to 24 g.

2) Cell line

3) drugs

Chlorogenic acid, coumaric quinic acid, chlorogenic acid: coumaric quinic acid (weight ratio 100:0.5), chlorogenic acid: coumaric quinic acid (weight ratio 100:0.1), chlorogenic acid : Coumaroyl quinic acid (weight ratio: 100:0.05), chlorogenic acid: coumaric quinic acid (weight ratio: 100:0.01), doxorubicin (ADM), physiological saline.

2 experimental methods

1) Cell culture

2) In vivo experiment

The ascites was taken from 7 to 10 days. After dilution, the number of cells was adjusted to (about 1×10 ⁶ cells), and the ascites cell suspension 0.2 ml·only ^{-1 was} inoculated subcutaneously in the left forelimb of the mouse. Animals were randomly divided into groups, 10 in each group, namely chlorogenic acid group, chlorogenic acid + coumaric quinic acid composition group, ADM group (2 mg·kg ^-1 ) and negative group (NS). Dosing was started on the second day after inoculation.

Chlorogenic acid group: intraperitoneal administration, once daily, administered at a dose of 30 mg·kg ^-1 .

The coumarinylquinic acid group was administered intraperitoneally once daily at a dose of 30 mg·kg ^-1 .

Chlorogenic acid + coumaric quinic acid composition group: intraperitoneal administration, once a day, the dose is 30 mg·kg ^-1 .

3 experimental results

See Table 3 and Figure 3.

Table 3 Effect of each experimental group on the growth volume of transplanted tumor of S180 sarcoma KM mice (

n=10)

Comparison of each drug group and negative group *p<0.05, **p<0.001

It can be seen from Table 3 and Figure 3:

1 The chlorogenic acid group, the chlorogenic acid + coumaroyl quinic acid group, and the ADM positive control group had significant inhibitory effects on the growth volume of S180 sarcoma KM mice at different time points during the experiment, and were negative. Compared with the control group, there were statistical differences;

2 The coumarinylquinic acid group had no significant inhibitory effect on the growth volume of transplanted tumor of S180 sarcoma KM mice;

3 chlorogenic acid + coumaric quinic acid group inhibited the growth volume of S180 sarcoma KM mice xenografts better than chlorogenic acid group and ADM positive control group;

See Table 4 and Figure 4.

Table 4 Effect of experimental groups on the weight and tumor inhibition rate of transplanted tumor of S180 sarcoma KM mice

Compared with the negative group ***p<0.001

It can be seen from Table 4 and Figure 4 that the inhibition rate of chlorogenic acid group, chlorogenic acid + coumaroyl quinic acid group and ADM positive control group on transplanted tumor of S180 sarcoma KM mice is higher than 45%; coumaroyl The inhibition rate of quinic acid group was the lowest, only 5%; the inhibition rate of chlorogenic acid + coumaric quinic acid group was the highest, indicating that the inhibition effect was the best, up to 64%.

At the same dose, the chlorogenic acid + coumaric quinic acid group was superior to chlorogenic acid and coumaric quinic acid alone, indicating that the combination of the two played a synergistic effect, among which chlorogenic acid and coumaric acid The weight ratio of quinic acid is preferably 100: (0.01 to 0.1), and still more preferably 100: 0.05.

The above experiment shows that the chlorogenic acid composition of the invention has obvious inhibitory effect on mouse sarcoma, and the inhibitory effect is better than that of doxorubicin (ADM), and the chlorogenic acid composition of the invention can be used for treating sarcoma with excellent curative effect. .

In summary, the chlorogenic acid of the present invention can effectively treat sarcoma, and the therapeutic effect is superior to the chemotherapy drug doxorubicin in the systemic administration + local administration mode, and it has been proved to be a safe drug, poison The side effects are very small, which can replace the traditional chemotherapy drugs for the treatment of sarcoma, which can effectively solve the problem that the traditional chemotherapy drugs have large side effects in the treatment of sarcoma, and the clinical application prospect is good.

Claims

Use of chlorogenic acid in the preparation of a medicament for treating sarcoma.
The use according to claim 1, characterized in that the medicament is prepared by using an effective amount of chlorogenic acid as an active ingredient, together with a pharmaceutically acceptable adjuvant or auxiliary ingredient.
The use according to claim 2, wherein the pharmaceutical preparation contains 1 to 3000 mg of chlorogenic acid per preparation unit.
The use according to claim 3, characterized in that the medicinal preparation has a chlorogenic acid used in an amount of from 1 to 100 mg/kg.
The use according to claim 3 or 4, characterized in that the medicament is an oral preparation or an injection.
The use according to claim 1, characterized in that the sarcoma comprises the sarcoma comprising osteosarcoma, fibrosarcoma, liposarcoma, rhabdomyosarcoma.
A pharmaceutical composition comprising chlorogenic acid and coumaric quinic acid.
The pharmaceutical composition according to claim 6, wherein the mass ratio of the chlorogenic acid to the coumaroyl quinic acid is from 100:0.01 to 0.5; preferably, the chlorogenic acid and coumaroyl quinone The mass ratio of nicotine is 100: 0.01 to 0.1; more preferably, the mass ratio of the chlorogenic acid to coumaroyl quinic acid is 100:0.05.
A method for preparing the pharmaceutical composition according to claim 7 or 8, characterized in that it is prepared by using chlorogenic acid and coumaric quinic acid as active ingredients, and adding pharmaceutically acceptable adjuvant to prepare a pharmaceutically acceptable drug. Formulation; Preferably, the preparation is an oral preparation or an injection preparation.
Use of the pharmaceutical composition of claim 7 or 8 in the manufacture of a medicament for the treatment of sarcoma.
The use according to claim 10, characterized in that the sarcoma comprises osteosarcoma, fibrosarcoma, liposarcoma, rhabdomyosarcoma.